Allergic asthma is a multi-faceted inflammatory disease in the lung. IgE and Th2-mediated inflammation play a critical role in extrinsic allergic asthma. Our labs have identified naturally processed IgE cytotoxic peptides (nECP) on IgE producing cells, which serve as CTL vaccine. The feasibility of using nECP with helper peptide PADRE as a universal IgE vaccine (or vaccine) to treat severe IgE-mediated allergic asthma is due to the following observations (preliminary results): (i) broad coverage of nECP supertype-based product concept for pan-IgE vaccine can be designed and applied to broad coverage of ethnically unbiased population. Second, progress was made for vaccine delivery platforms, e.g., transcutaneous immunization with nECP/PADRE in an FDA approved imiquimod for topical use, followed by nECP/PADRE in saline intranasally (IN). The mucosal immunization stimulating dendritic cells (DCs), and CD8 T cells enriched the lung to protect against IgE production in the rodent model. (iii) safety of the vaccine in its unique IgE isotype suppression, while sparing other antibody isotypes; and IgE recovery after the treatment window. (iv) testing human supertype nECP in the HLA-A, -B tg rodents, and in particular testing IgE suppression by in vitro nECP CTL immunization of human PBMCs, which suppress human IgE production. The goal is to develop a commercial product of supertype based universal IgE peptide vaccine with broad HLA-A,-B supertype coverage to treat IgE-mediated severe asthma. The vaccine will suppress human IgE producing B cells/plasma cells, including long-lived plasma cells in human lung in a safe IgE isotype-specific therapeutic window, permitting IgE recovery and parasite defense. The top priority is to compete studies in the rodent model and supertype composition of human nECP vaccine. The next Development Phase will require venture capital, the pharmas capital or licensing revenue, which is dependent on the completion of Research Phase of the three Aims proposed through this SBIR grant application for CMC or cGMP of vaccine manufacture. Our three aims are as follow: Aim 1. Evaluate efficacy and safety of a universal nECP asthma vaccine in the rodent model Aim 2. Evaluate the routes and mechanisms of therapeutic vaccination Aim 3. Evaluate a universal human nECP supertype asthma vaccine for 86% population coverage Milestones Aim 1 Aim 2 Aim 3 Year 1 Protection and recovery cycle; evaluating different helper peptides for sustaining protective CD 8 T cell responses; tolerant/unresponsive CD 8 T cells and restore responsiveness/breaking tolerance. The role of cDCs in the skin via TCI vaccination by FACS and analysis of migratory cDCs by T cell stimulation; the role cDCs via NALT and nasal tract in the B6 model; function of cDCs excluding blood monocytes. Evaluate high affinity human nECPs in A3 and more B7, A2 supertypes in inhibiting canonical peptide binding to purified HLA-A, -B. Year 2 Analysis of changes in B- cells and IgE plasma cells following CTL attack; IgE clearance; changes in submucosal mast cells, IgE/ mast cell axis after TCI/IN vaccination; AHR and Airway Remodeling analysis. Analysis of langerin+ DTR knockout of migratory CD103+ cDCs; Analysis of TCM, TEM and TRM In lymphoid and nonlymphoid organs after TCI or IN or TCI/IN, longevity of each subset with/without PADRE; in vivo conversion of TEM vs. TRM; the role of TCM. Tg studies for A3 supertypes in A3Kb tg mice, and more B7 and A2 supertypes in B7, A2 tg mice; high resolution mapping in human PBMC (also cryopreserved for a large aliquots) for combined supertype epitopes for vitro CTL for IgE suppression in vitro. Year 3 Confirm protection in airway inflammation and AHR in the Aspergillus fumigatus and cockroach models using TCI/IN vaccination. Analysis of trafficking in langerin+ CD103+ cDCs depletion alone vs. combined CD103+ cDCs and CD103+ TRM in CD103 knockout mice. Investigating the strength/magnitude by IFN-? spots and lytic potential and quality of CTL of multiple supertype induced cultures, and expand into three additional supertypes: A1, A24 and B44 for high percentage human coverage.